Differential Labeling of Glycoproteins with Alkynyl Fucose Analogs.
Differential Labeling of Glycoproteins with Alkynyl Fucose Analogs.
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DOI：
10.3390/ijms21176007
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发表时间：
2020-08-20
影响因子：
5.6
通讯作者：
Kizuka Y
中科院分区：
文献类型：
--
作者：
Ma C;Takeuchi H;Hao H;Yonekawa C;Nakajima K;Nagae M;Okajima T;Haltiwanger RS;Kizuka Y
岩藻糖基化聚糖对蛋白质和细胞的生理功能有重要的调节作用。岩藻糖基化聚糖水平的改变与各种疾病相关。对于岩藻糖基化聚糖的检测和功能调节，使用岩藻糖（Fuc）类似物的化学生物学方法是有用的。然而，关于在岩藻糖基化聚糖的生物合成途径中酶如何有效地利用每个非天然Fuc类似物知之甚少。我们在这里展示了三种具有相似但不同结构的可点击Fuc类似物以不同的效率和蛋白质特异性标记细胞聚糖。例如，6-炔基（Alk）-Fuc修饰的O-Fuc聚糖比7-Alk-Fuc更有效。细胞中产生的GDP-6-Alk-Fuc水平也高于GDP-7-Alk-Fuc水平。全面的体外岩藻糖基转移酶测定显示，7-Alk-Fuc通常被大多数岩藻糖基转移酶耐受。令人惊讶的是，两种蛋白质O-岩藻糖基转移酶（POFUT）可以在体外转移所有Fuc类似物，可能是因为POFUT结构在其Fuc结合位点周围具有更大的空间。这些发现表明，用Fuc类似物标记和检测岩藻糖基化聚糖取决于多个细胞步骤，包括转化为GDP形式，转运到ER或高尔基体中，以及被每种岩藻糖基转移酶利用，从而为设计新型糖类似物以特异性检测靶聚糖或抑制其功能提供了见解。
Fucosylated glycans critically regulate the physiological functions of proteins and cells. Alterations in levels of fucosylated glycans are associated with various diseases. For detection and functional modulation of fucosylated glycans, chemical biology approaches using fucose (Fuc) analogs are useful. However, little is known about how efficiently each unnatural Fuc analog is utilized by enzymes in the biosynthetic pathway of fucosylated glycans. We show here that three clickable Fuc analogs with similar but distinct structures labeled cellular glycans with different efficiency and protein specificity. For instance, 6-alkynyl (Alk)-Fuc modified O-Fuc glycans much more efficiently than 7-Alk-Fuc. The level of GDP-6-Alk-Fuc produced in cells was also higher than that of GDP-7-Alk-Fuc. Comprehensive in vitro fucosyltransferase assays revealed that 7-Alk-Fuc is commonly tolerated by most fucosyltransferases. Surprisingly, both protein O-fucosyltransferases (POFUTs) could transfer all Fuc analogs in vitro, likely because POFUT structures have a larger space around their Fuc binding sites. These findings demonstrate that labeling and detection of fucosylated glycans with Fuc analogs depend on multiple cellular steps, including conversion to GDP form, transport into the ER or Golgi, and utilization by each fucosyltransferase, providing insights into design of novel sugar analogs for specific detection of target glycans or inhibition of their functions.
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影响因子：
16.6
作者：
Chang, Pamela V.;Chen, Xing;Smyrniotis, Chris;Xenakis, Alexander;Hu, Tianshun;Bertozzi, Carolyn R.;Wu, Peng
通讯作者：
Wu, Peng
影响因子：
11.1
作者：
Kizuka Y;Kitazume S;Fujinawa R;Saito T;Iwata N;Saido TC;Nakano M;Yamaguchi Y;Hashimoto Y;Staufenbiel M;Hatsuta H;Murayama S;Manya H;Endo T;Taniguchi N
通讯作者：
Taniguchi N
DOI：
10.1073/pnas.0811481106
发表时间：
2009-01-06
影响因子：
11.1
作者：
Laughlin, Scott T.;Bertozzi, Carolyn R.
通讯作者：
Bertozzi, Carolyn R.
影响因子：
4.8
作者：
Jarva, Michael A.;Dramicanin, Marija;Goddard-Borger, Ethan D.
通讯作者：
Goddard-Borger, Ethan D.
影响因子：
2.9
作者：
Ito, Jun;Herter, Thomas;Heazlewood, Joshua L.
通讯作者：
Heazlewood, Joshua L.